nm23-H_1基因转染前后人高转移大细胞肺癌细胞株L9981 PKA活性变化的实验研究  

Experimental study of effects of nm23-H_1 gene transfection on the change of PKA activity in human high-metastasis large cell lung cancer cell line L9981

作　　者：李定彪[1] 周清华[1] 王艳萍[1] 朱文[1] 陈小禾[1] 杨俊杰[1] 刘伦旭[1] 付军科[1] 聂强[1] 李印[1] 

机构地区：[1]四川大学华西医院四川省肺癌分子重点实验室,胸外科,成都610041

出　　处：《中国肺癌杂志》2004年第2期91-94,共4页Chinese Journal of Lung Cancer

基　　金：国家自然科学基金 (No .30 0 70 333和No.30 1 0 0 0 75)资助~~

摘　　要：目的　探讨nm2 3 H1基因转染和forskolin对人高转移大细胞肺癌细胞株L9981PKA活性的影响。方法　应用PKA通路特异激动剂forskolin对人高转移大细胞肺癌原代细胞株L9981、转基因细胞株L9981 nm 2 3 H1 pLXSN和空载体转染细胞株L9981 pLXSN共同培养 ,应用放免法检测forskolin处理后不同时间点三个细胞株PKA的活性变化。结果　 ( 1)forskolin处理前L9981、L9981 pLXSN和L9981 nm2 3 H1 pLXSN的PKA活性经F检验有显著性差异 (P <0 .0 1) ;两两比较 :L9981 nm2 3 H1 pLXSN的PKA活性显著高于L9981和L9981 pLXSN(P <0 .0 1) ,而L9981与L9981 pLXSN之间比较无显著性差异 (P >0 .0 5 )。 ( 2 )在同一作用时间 ( 3 0min)下 ,三个细胞株经不同浓度forskolin处理后PKA活性均显著升高 (P <0 .0 1) ,在forskolin浓度为 10 0 μmol/L时PKA活性最高 ,呈剂量依赖关系。 ( 3 )三个细胞株在同一浓度forskolin( 10 0μmol/L)处理下 ,PKA活性随作用时间升高 ,在forskolin作用时间为 3 0min时PKA的活性最高 ,呈时间依赖关系。结论　 ( 1)nm2 3 H1基因转染能显著上调人高转移大细胞肺癌细胞株L9981的PKA活性 ,nm2 3 H1作为一种肿瘤转移抑制基因的作用机理与PKA信号通路可能有一定联系 ;( 2 )forskolin能显著上调L9981细胞?Objective To explore the effects of nm23-H1 gene transfection and forskolin on PKA activity in human high-metastasis large cell lung cancer cell line L9981. Methods Three cell lines, primary human large cell lung cancer cell line (L9981), vector transfection cell line (L9981-pLXSN) and nm23-H1 gene transfection cell line (L9981-nm23-H1-pLXSN), were treated with PKA activator forskolin. The PKA activity at different time points after treatment with forskolin was detected in the three lung cancer cell lines by radioimmunological method with SignaTECT cAMP-dependent PKA assay system. Results (1) Before forskolin treatment, the activity of PKA of L9981-nm23-H1-pLXSN was remarkably higher than those of L9981 and L9981-pLXSN (P<0.01), but no significant difference in the PKA activity was observed between L9981 and L9981-pLXSN (P>0.05). (2)The PKA activity was remarkably increased in all the three lung cancer cell lines after treatment with different concentration of forskolin (P<0.01), and up to the highest level at the concentration of 100 μmol/L. It showed a dose-dependent relationship between the PKA activity and forskolin concentration; (3) The PKA activity in all the three cell lines was elevated to the highest level at 30 minutes after treatment with forskolin of 100 μmol/L, and it showed a time-dependent relationship between the PKA activity and action time of forskolin. Conclusion (1)Transfection of nm23-H1 gene can up-regulate the PKA activity of human high-metastasis large cell lung cancer cell line L9981, and its function as a tumor metastasis suppressor gene may be related to its effects on regulation of PKA signal transduction pathways; (2)Forskolin can remarkably up-regulate the PKA activity of L9981 cell line, and the elevation of PKA activity has a time-dependent and dose-dependent relation to forskolin.

关 键 词：NM23-H1 基因转染 癌细胞 L9981 PKA活性 实验 FORSKOLIN PKA通路特异激动剂 高转移 大细胞肺癌 

分 类 号：R734.2[医药卫生—肿瘤] R394[医药卫生—临床医学]